Ultrasensitive sniff-cam for biofluorometric-imaging of breath ethanol caused by metabolism of intestinal flora

Kenta Iitani, Koji Toma, Takahiro Arakawa, Kohji Mitsubayashi

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

We developed a gas-imaging system (sniff-cam) for gaseous ethanol (EtOH) with improved sensitivity. The sniff-cam was applied to measure the extremely low concentration distribution of breath EtOH without the consumption of alcohol, which is related to the activity of the oral or gut bacterial flora. A ring-type ultraviolet-light-emitting diode was mounted around a camera lens as an excitation light source, which enabled simultaneous excitation and imaging of the fluorescence. In the EtOH sniff-cam, a nicotinamide adenine dinucleotide (NAD)-dependent alcohol dehydrogenase (ADH) was used to catalyze the redox reaction between EtOH and the oxidized form of NAD (NAD+). Upon application of gaseous EtOH to the ADH-immobilized mesh that was soaked in an NAD+ solution and placed in front of the camera, NADH was produced through an ADH-mediated reaction. NADH expresses fluorescence at an emission wavelength of 490 nm and excitation wavelength of 340 nm. Thus, the concentration distribution of EtOH was visualized by measuring the distribution of the fluorescence light intensity from NADH on the ADH-immobilized mesh surface. First, a comparison of image analysis methods based on the red-green-blue color (RGB) images and the optimization of the buffer pH and NAD+ solution concentration was performed. The new sniff-cam showed a 25-fold greater sensitivity and broader dynamic range (20.6-300000 ppb) in comparison to those of the previously fabricated sniff-cam. Finally, we measured the concentration distribution of breath EtOH without alcohol consumption using the improved sniff-cam and obtained a value of 116.2 ± 35.7 ppb (n = 10).

Original languageEnglish
Pages (from-to)9458-9465
Number of pages8
JournalAnalytical Chemistry
Volume91
Issue number15
DOIs
Publication statusPublished - 2019 Aug 6
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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